RRC ID 44019
Author Liang H, Baudouin C, Daull P, Garrigue JS, Brignole-Baudouin F.
Title Ocular safety of cationic emulsion of cyclosporine in an in vitro corneal wound-healing model and an acute in vivo rabbit model.
Journal Mol. Vis.
Abstract PURPOSE:Topical preparations of cyclosporine (CsA) are common therapeutics for the treatment of dry eye. However, they are not devoid of side effects, such as allergy and irritation. The present study aimed at evaluating the safety profile of a new CsA formulation in cationic emulsion (CEm) in vitro with a dynamic corneal wound healing assay using human corneal epithelial (HCE) cells, and in vivo in a rabbit acute toxicity model.
METHODS:Three different csa formulations were tested: 1) 0.05%CsA-CEm, 2) commercial 0.05%CsA-Anionic emulsion (CsA-AEm, Restasis®), and 3) 0.05%CsA-Oil solution. Phosphate buffered saline (PBS) was used as negative control and 0.02% benzalkonium chloride (BAK) as the toxic control. In vitro, a wound was created by scratching through a confluent HCE cell layer and exposed 30 min to 1/10 dilutions of the different formulations. Cytotoxicity, cell migration, and proliferation were performed to analyze the recovery at days 1, 2, and 3. In vivo, the eye drops were applied to rabbit eyes 15 times at 5-min intervals. The ocular surface structures were examined with a slit-lamp and by corneal in vivo confocal microscopy (IVCM) for detailed examination of corneal epithelium, stroma, limbus, and conjunctiva-associated lymphoid tissue (CALT) structures.
RESULTS:The in vitro study confirmed that a 0.02% BAK solution delayed the corneal healing process (-57%) by severely damaging the remaining HCE cells. The other formulations maintained a normal healing rate with a similar behavior for CsA-CEm, CsA-AEm, and PBS with no significant differences (at D3, 66%-74% closure). In the rabbit, 0.02%BAK showed the highest toxicity, inducing redness, chemosis with damaged corneal epithelium, and inflammatory cell infiltrations. CsA-AEm and CsA-Oil induced moderate infiltrations of inflammatory cells around the CALT. CsA-CEm presented the lowest toxicity with patterns similar to PBS.
CONCLUSIONS:The combination of these in vitro and in vivo models evaluated the tolerance/cytotoxicity and the dynamic wound healing potential of CsA in different formulations. While CsA-AEm, CsA-CEm, and CsA-Oil are generally well tolerated, only CsA-CEm appeared to maintain the HCE cells' normal healing rate in vitro and low levels of inflammation in vivo.
Volume 18
Pages 2195-204
Published 2012
PMID 22919267
PMC PMC3425577
MeSH Animals Benzalkonium Compounds / administration & dosage Benzalkonium Compounds / adverse effects Cell Movement / drug effects Cell Proliferation / drug effects Cell Survival / drug effects Conjunctiva / drug effects* Cornea / cytology Cornea / drug effects* Cyclosporine / pharmacology* Emulsions Epithelial Cells / cytology Epithelial Cells / drug effects* Epithelium, Corneal / cytology Epithelium, Corneal / drug effects* Humans Microscopy, Confocal Models, Biological Ophthalmic Solutions / administration & dosage Preservatives, Pharmaceutical / administration & dosage Preservatives, Pharmaceutical / adverse effects Rabbits Wound Healing / drug effects
IF 2.219
Times Cited 16
WOS Category OPHTHALMOLOGY BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells